Hematological profile of healthy Thoroughbred foals from birth to one month of age in Trinidad, West Indies.

OBJECTIVE: This study aimed to develop a sound database for the hematological reference intervals of thoroughbred foals in Trinidad, West Indies from birth to 1 month of age. ANIMALS: 89 foals. METHODS: Whole blood samples were taken from 89 foals throughout Trinidad at approximately 1 day, 1 week, and 1 month of age. These foals were examined to be classified as healthy or free from disease. Complete blood count (CBC), microscopic analysis of blood smears, and conventional PCR for Theileria equi and Babesia caballi were performed. RESULTS: Of the 89 foals, 67 were deemed healthy and suitable for establishing reference intervals. Foals in this study had lower mean hemoglobin and hematocrit values for all 3 times of sampling when compared to their North American counterparts. Age had a significant effect on hemoglobin, hematocrit, white blood cell (WBC), neutrophil, and platelet counts of the foals from birth to 1 month of age. CLINICAL RELEVANCE: Variations in reference intervals can occur due to differences in demographic, physiological, and environmental factors such as age, gender, breed, and geographical location. Given the changes in the hematological values over time, this study provides clinicians with valuable information that can be used to monitor the health status of newborn foals and detect disease conditions.

Completing the Puzzle: A Cluster of Hunting Dogs with Tick-Borne Illness from a Fishing Community in Tobago, West Indies.

Eight hunting dogs were visited by a state veterinarian on the island of Tobago, Trinidad and Tobago, West Indies, as owners reported anorexia and paralysis in five of their dogs. The veterinarian observed a combination of clinical signs consistent with tick-borne illness, including fever, anorexia, anaemia, lethargy and paralysis. Blood and ticks were collected from each dog and submitted to a diagnostic laboratory for analysis. Microscopic analysis revealed a mixed infection of intracytoplasmic organisms consistent with Babesia spp. (erythrocyte) and Ehrlichia spp. (monocyte), respectively, from one dog, while a complete blood count indicated a regenerative anaemia (n = 1; 12.5%), non-regenerative anaemia (n = 4; 50%), neutrophilia (n = 3; 37.5%), lymphocytosis (n = 2; 25%), thrombocytopaenia (n = 3; 37.5%) and pancytopaenia (n = 1; 12.5%). DNA isolated from the eight blood samples and 20 ticks (16 Rhipicephalus sanguineus and 4 Amblyomma ovale) were subjected to conventional PCR and next-generation sequencing of the 16S rRNA and 18S rRNA gene for Anaplasma/Ehrlichia and Babesia/Theileria/Hepatozoon, respectively. The DNA of Ehrlichia spp., closely related to Ehrlichia canis, was detected in the blood of three dogs (37.5%), Anaplasma spp., closely related to Anaplasma marginale, in two (25%), Babesia vogeli in one dog (12.5%) and seven ticks (35%) and Hepatozoon canis and Anaplasma spp., in one tick (5%), respectively. These findings highlight the need to test both the vector and host for the presence of tick-borne pathogens when undertaking diagnostic investigations. Further studies are also warranted to elucidate the susceptibility of canids to Anaplasma marginale.

First reported case of leishmaniasis in a cat in Trinidad and Tobago.

A 3-year-old, female, domestic shorthair cat, was presented to the Veterinary Teaching Hospital at the School of Veterinary Medicine (SVM), Trinidad and Tobago for a swollen nose, and multiple, variably sized small masses on both ears. The initial diagnostic tests included a CBC, serum biochemistry profile, cytological evaluation of masses on the ear and nose, and FeLV/FIV testing. The CBC and biochemistry results were unremarkable except for a hyperproteinaemia and hyperglobulinemia. Cytology of the nose and ear lesions revealed mixed inflammation and high numbers of intracellular and extracellular organisms consistent with Leishmania amastigotes. The cat was FeLV/FIV negative. Histopathology and Leishmania IFA and PCR analysis were subsequently performed, confirming the Leishmania diagnosis. The PCR, DNA sequencing and phylogenetic tree analyses identified L. amazonensis. This is the first reported case of L. amazonensis infection in a domestic animal in Trinidad with molecular characterization indicating it exists in the region and is likely being transmitted by sandflies.

Acute and subacute toxicity evaluation of hydroalcoholic extract from the stem bark of Bois Bande (Parinari campestris Aubl.1772) in rats.

INTRODUCTION: The bark of Bois Bande (Parinari campestris) is a popular aphrodisiac in the Caribbean that has been traditionally used for many years to restore sexual vitality, increase sperm count, and treat erectile dysfunction, without valid scientific data. Acute and 28-day subacute toxicity studies were conducted to evaluate the safety of the hydroalcoholic extract of P.campestris bark and to find a safe dose for human use in conventional medicine. METHODS: The acute toxicity study used a single oral dose of P.campestris extract at four separate doses, 5, 50, 300, and 2,000 mg/kg, and was seen for 14 days, while the subacute toxicity study used a daily oral dose of P.campestris extract at 3 different doses, 100, 300, and 1000 mg/kg/day for 28 days. RESULTS: The LD50 of P.campestris extract was found to be greater than 2000 mg/kg in the acute toxicity study. P.campestris extract did not show toxicity at 1000 mg/kg/day in subacute toxicity trial; NOAEL was 1000 mg/kg/day in rats. However, the body weight was increased in males. CONCLUSION: In conclusion, 1000 mg/kg P.campestris extract can be considered safe and non-toxic in males.

Disseminated Mycobacterium avium subsp. hominissuis infection and ascites in an FIV-positive cat.

A domestic shorthair cat was presented to the Veterinary Teaching Hospital at The University of the West Indies with a history of anorexia, ataxia, and lethargy. On physical examination, moderate abdominal distension and a palpable abdominal fluid wave were noted. Dark yellow, cloudy fluid was collected via abdominocentesis. Fluid analysis indicated that the effusion was a transudate containing low numbers of macrophages and occasional neutrophils. Some of the macrophages contained rod-shaped nonstaining structures of variable length (2-4 um). These structures were also seen extracellularly in low numbers. The morphology of the structures was suggestive of Mycobacterium. The cat's condition continued to deteriorate, and it died within a few hours of being admitted. Further diagnostic tests revealed feline immunodeficiency virus (FIV) infection with concurrent Mycobacterium avium subsp hominissuis infection. To the authors' knowledge, this is the first reported case of nontubercular mycobacterial-related ascites in a cat.

Granulocytopenia associated with thymoma in a domestic shorthaired cat.

A 5-year-old, spayed female cat was referred because of a mass in the cranial mediastinum noted on thoracic radiographs. A thymoma was diagnosed following ultrasound and biopsy of the mass. Treatment was initiated with coarse-fraction radiation therapy using external-beam therapy (four fractions of 5 Gy). The mass responded, but granulocytopenia developed. Bone marrow examination showed a myeloid to erythroid ratio of approximately 1:1, with a left shift within the myeloid line. These findings, as well as the lack of toxic changes within the peripheral blood neutrophils, suggested immune-mediated destruction of peripheral granulocytes. Immune suppression with prednisone and cyclosporine was instituted. After 7 weeks, the neutrophil count returned to normal. The tumor was removed, and cyclosporine was reduced and eventually discontinued 3 weeks postsurgery.

A Multicentric T-Cell Lymphoma with a Plasmacytoid Morphology in a Dog.

An 8-year-old male (neutered) Labrador with a history of erythematous skin lesions and exercise intolerance for a prolonged period was suddenly found dead. Necropsy findings revealed an infiltrative, focally extensive mass which occupied 25% of the cardiac interventricular septum. Severe endocardiosis was also found on the bicuspid and tricuspid valves. The submandibular lymph nodes and kidneys were bilaterally enlarged, and the pre-hepatic lymph node and spleen were also enlarged. Multiple dermal pustules were present around the mouth and on the ear, and small ulcers were present on the tongue. Histopathological examination detected the presence of neoplastic lymphocytes with a plasmacytoid morphology in these tissues as well as in the tongue and skin lesions. Immunohistochemical (CD3⁺/CD18⁺) evaluation was consistent with a T-cell lymphoma, which could be classified as a peripheral T-cell lymphoma, not otherwise specified (PTCL-NOS).

A Case Report of Disseminated Canine Histiocytic Sarcoma in Trinidad and Tobago.

Ocular histiocytic sarcomas (as a presenting part of disseminated histiocytic sarcoma) are not commonly diagnosed. A 10-year-old female intact Rottweiler presented to the School of Veterinary Medicine, Trinidad with buphthalmia and pain in the left eye. The cornea of the left eye appeared diffusely opaque with a conjunctival mucopurulent ocular discharge. A thorough ophthalmic assessment identified an intraocular proliferative tumor to which a unilateral enucleation was performed, however the animal died soon after. Post mortem examination and light microscopy revealed that the intraocular lesion with visceral macro-metastases was in fact a histiocytic sarcoma. Further to this, immune-phenotyping was performed to confirm the diagnosis of disseminated histiocytic sarcoma. This is the first time such a tumor has been diagnosed in Trinidad and Tobago.

Oral Plasmacytoma in a Dog.

A 6-year-old male Pit bull mix dog presented for bleeding from the mouth persisting for five days. A clinical evaluation revealed a 2 × 3 cm soft tissue mandibular mass at the crown of the first premolar, as well as a non-regenerative anemia and hyperproteinemia. Cytologic and histopathologic evaluations of the mass were compatible with an oral plasmacytoma.

Prospective study investigating transplacental transmission of equine piroplasmosis in thoroughbred foals in Trinidad.

Equine piroplasmosis caused by Theileria equi and Babesia caballi is endemic in Trinidad and Tobago. Transmission occurs by ticks of the family Ixodidae. T. equi can also be transmitted transplacentally; however transplacental transmission of B. caballi is unknown. This study aims to investigate transplacental transmission of equine piroplasmosis from thoroughbred mares naturally infected via the tick vector. Whole blood and serum samples were collected from 117 mares in the fifth month of pregnancy. Blood samples were also collected from each of their foals (89 in total) within the first 36h of birth. Additionally, all foals were observed for clinical signs within 30days post - partum. All samples were examined microscopically for intra-erythrocytic piroplasms. Serum ELISA tests and PCR analysis on whole blood were performed to determine the presence of T. equi and B. caballi. Thirty-four (30.6%) mares and 14 (15.7%) of their foals were seropositive for T. equi. Twenty-seven (24.3%) mares were positive for T. equi by conventional (c) PCR. Real time (q) PCR analysis based on the ema - 1 gene revealed that seven (8%) foals were positive for T. equi. Eighty-nine (76.1%) mares and 38 (42.7%) foals were seropositive for B. caballi. Four (3.4%) mares were positive for B. caballi by cPCR. Three out of the four cPCR positive mares either had resorptions, or stillbirths for that pregnancy. From this study, there is strong evidence that transplacental transmission of B. caballi can occur leading to foetal losses. Six foals (7%) were positive for B. caballi by qPCR. Of these six, four were born to B. caballi seropositive mares. In this study a foal born of a T. equi seropositive mare was 55.7 times more likely to be serologically positive for T. equi than a foal born to a T. equi seronegative mare. Similarly a foal born of a B. caballi seropositive mare was 39.4 times more likely to be serologically positive for B. caballi than a foal born to a mare that was serologically negative for B. caballi at the fifth month of pregnancy. This is as a result of the ingestion of colostrum containing antibodies to these pathogens. Mares should be screened during pregnancy and their foals closely monitored at parturition for evidence of equine piroplasmosis so that treatment can be implemented earlier for a better prognosis.

Haematological parameters in stray dogs seropositive and seronegative to Ehrlichia canis in North Trinidad.

In view of the fact that stray dogs are a reservoir for many diseases, this study was undertaken to determine the prevalence of Ehrlichia canis in stray dogs in North Trinidad and to evaluate the diagnostic implications of haematological alterations associated with seropositivity. Overall, 41 (44.6%) of 92 stray dogs were seropositive to E. canis by the indirect immunofluorescent antibody test. Dogs, one year of age and older (59.7%) were more likely to be seropositive than dogs less than one year old (13.3%) (p<0.001). No significant differences in seropositivity between females and males were found. The odds ratios showed that seropositive dogs were 3.34 (CI 95%; 1.33-8.59) and 5.17 (CI 95%; 0.19-1.26) times more likely to have low platelet counts and elevated total serum protein concentrations (p=0.014 and p<0.001, respectively) than seronegative dogs. Lower mean platelet counts and a higher mean total protein concentration were associated with seropositivity (p<0.01). Mean eosinophil and segmented neutrophil counts were elevated in dogs that tested negative for E. canis antibodies (p=0.002 and p<0.005, respectively). Other haematological parameters were not different between the 2 groups. The high percentage of stray dogs infected with E. canis should alert veterinarians to the potential risk of transmission of the disease. A comprehensive study possibly using molecular methods such as nested PCR should be undertaken to determine how co-infection with other pathogens may alter haematological profiles. In general, control of ticks and stray dog populations may help to control the spread of tick-borne diseases.

A case of transplacental transmission of Theileria equi in a foal in Trinidad.

Equine piroplasmosis due to Theileria equi and Babesia caballi is endemic in Trinidad. A case of equine piroplasmosis due to T. equi was diagnosed in a thoroughbred foal at 10h post-partum. A high parasitaemia (63%) of piroplasms was observed in a Wright-Giemsa(®) stained thin blood smear from the foal. In addition, the 18S rRNA gene for Babesia/Theileria was amplified from DNA extracted from the blood of the foal and the mare. Amplified products were subjected to a reverse line blot hybridization assay (RLB), which confirmed the presence of T. equi DNA in the foal. The mare was negative by RLB but was positive for T. equi using a nested PCR and sequence analysis. In areas where equine piroplasmosis is endemic, severe jaundice in a post-partum foal may be easily misdiagnosed as neonatal isoerythrolysis. Foals with post-partum jaundice should be screened for equine piroplasmosis, which may be confirmed using molecular methods if available.

The influence of age and Rhodococcus equi infection on CD1 expression by equine antigen presenting cells.

UNLABELLED: There is a distinct age-associated susceptibility of horses to Rhodococcus equi infection. Initial infection is thought to occur in the neonatal and perinatal period, and only foals less than 6 months of age are typically affected. R. equi is closely related and structurally similar to Mycobacterium tuberculosis, and causes similar pathologic lesions. Protective immune responses to M. tuberculosis involve classical major histocompatibility complex (MHC)-restricted T cells that recognize peptide antigen, as well as MHC-independent T cells that recognize mycobacterial lipid antigen presented by CD1 molecules. Given the structural similarity between these two pathogens and our previous observations regarding R. equi-specific, MHC-unrestricted cytotoxic T lymphocytes (CTL), we developed 3 related hypotheses: (1) CD1 molecules are expressed on equine antigen presenting cells (APC), (2) CD1 expression on APC is less in foals compared to adults and (3) infection with live virulent R. equi induces up-regulation of CD1 on both adult and perinatal APC. CD1 expression was examined by flow cytometric analysis using a panel of monoclonal CD1 antibodies with different species and isoform specificities. RESULTS: Three CD1 antibodies specific for CD1b showed consistent cross reactivity with both foal and adult monocyte-derived macrophages (MDM). CD1b and MHC class II expression were significantly higher on adult MDM compared with foals. R. equi infected MDM showed significantly lower expression of CD1b, suggesting that infection with this bacterium induces down-regulation of CD1b on the cell surface. Histograms from dual antibody staining of peripheral blood mononuclear cells also revealed that 45-71% of the monocyte population stained positive for CD1b, and that the majority of these also co-expressed MHC II molecules, indicating that they were APC. The anti-CD1 antibodies showed no binding or minimal binding to bronchoalveolar lavage (BAL)-derived macrophages. CONCLUSION: The CD1b isoform is evolutionarily conserved, and is present on equine MDM, as well as on circulating blood monocytes. The unique susceptibility of foals to R. equi infection may be due in part to lower expression of CD1 and MHC class II, as observed in this study. The data also suggests that infection with R. equi induces down-regulation of CD1b on equine MDM. This may represent a novel mechanism by R. equi to avoid detection and killing of infected cells by the immune system, similar to that observed when human APC are infected with M. tuberculosis.